Radioimmunoassay of Prostaglandins Fα, E1 and E2 in Human Plasma |
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Authors: | F Dray B Charbonnel J Maclouf |
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Institution: | Unitéde Radioimmunologie Analytique, Institut Pasteur, 28 rue du Dr. Roux, Paris, France |
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Abstract: | Abstract. Antibodies against prostaglandins (PG)F2α, E1 and E2 were obtained in rabbits immunized with respectively PG F2α, PG E1 and PG E2 conjugated to bovine serum albumin by carbodiimide. A radioimmunoassay capable of measuring 7 pg of PG Fα, 2 pg of PG E2 and 14 pg of PG Ej in human peripheral plasma is described. Plasma samples (pH 3, citric acid) are extracted with cyclohexane: ethyl acetate, 1:1 and then chromatographed on silicic acid columns to separate the prostaglandins into three fractions: fraction I, PG A, PG B and some unknown immunoraactive compounds; fraction II, PG E and fraction III PG Fα. The recovery is 80 %± 6. 2. Mean plasma levels iu adults of PG Fa and PG E, expressed in pg/ml: -PG Fα 12 ± 2. 8 (n = 25 men), 8 ± 2. 3 (n = 18 women, follicular phase), 7 ± 1. 4 (n = 18 women, luteal phase). -PG E1 40. 5 + 7. 6 (n = 13 men), 38 + 17. 1 (n = 10 women). -PG E2 4. 5 ± 1 (n = 12 adult subjects). The major characteristics of the method described herein are the following: - a large volume of plasma has to be processed (10 ml or more for PG Fa and PG E1, 5 ml or more for PG E2). - a chromatographic step is necessary to separate the different prostaglandins which makes it possible to circumvent problems of immunological cross reactivity and interference with unknown immunoreactive compounds. - great care has been taken in collection of blood samples, especially to insure complete removal of blood cells namely platelets. |
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Keywords: | Prostaglandins radioimmunoassay in human plasma |
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